急性痛风性关节炎模型大鼠炎性反应及Toll样受体通路相关蛋白表达  

Inflammatory pathways and Toll-like receptors related protein expression on acute gouty arthritis model rats

作　　者：刘剑刚[1] 杨钊田 杨卫彬[3] Liu Jiangang;Yang Zhaotian;Yang Weibing(Xiyuan Hospital of China Academy of Chinese Medicine Sciences,Ceremony of National Clinical Research Center for Chinese Medicine Cardiology,Beijing 100091,China;Rheumatoid Immune Onset of Shaanxi University of Traditional Chinese Medicine Hospital,Xian 712000,China;Graduate School of China Academy of Chinese Medicine Sciences,Beijing 100700,China)

机构地区：[1]中国中医科学院西苑医院心血管病中心国家中医心血管病临床医学研究中心,北京100091 [2]陕西中医药大学附属医院风湿免疫血液科,西安712000 [3]中国中医科学院研究生院,北京100700

出　　处：《中华风湿病学杂志》2021年第11期747-753,I0004,共8页Chinese Journal of Rheumatology

基　　金：北京市科学技术委员会G20工程创新研究项目(Z17110000171706)。

摘　　要：目的采用尿酸钠诱导实验性大鼠急性痛风性关节炎模型,观察大鼠的炎性反应及对踝关节Toll样受体相关蛋白表达和双氯芬酸钠的干预作用。方法雄性无特定病原体级Wistar大鼠30只,随机数字表法将大鼠分为空白对照组(0.9%氯化钠注射液组)10只、模型组10只、双氯芬酸钠缓释片药物组(双氯芬酸钠片组,1.35 mg/kg体质量)10只。将尿酸钠晶体充分研磨后,加入适量0.9%氯化钠注射液和吐温-80(9∶1)制成混悬液进行造模,空白对照组大鼠同样位置注射0.9%氯化钠注射液。于造模后开始灌胃给药,0.9%氯化钠注射液组和模型组给予等容积纯净水,每天1次,共7 d,采用足趾容积装置测定大鼠(造模后4、8、24、48、72 h)的关节肿胀度,麻醉后腹主动脉取血测定大鼠肾功能、IL-1β、IL-6和TNF-α含量,采用蛋白质免疫印迹法和免疫组织化学法测定大鼠踝关节Toll样受体4(TLR4)、髓样分化因子88(MyD88)、NF-κBp65的蛋白表达。多组比较采用单因素方差分析,2组间比较采用LSD-t法,不同时间段的比较采用重复测量的方差分析法。结果造模后的大鼠右踝关节出现不同程度的红肿、行走迟缓、反应迟钝等症状。光学镜下观察,模型组大鼠踝关节滑膜细胞增生,局部可见细胞变性坏死和炎性细胞浸润。和0.9%氯化钠注射液组IL-1β、IL-6和TNF-α[(24.6±2.6)pg/ml,(132±20)pg/ml和(72±6)pg/ml]比较,模型组[(28.4±4.3)pg/ml,(173±26)pg/ml和(81±5)pg/ml]升高(t=2.601,P<0.05;t=3.375,P<0.01;t=1.392,P>0.05);和模型组比较,双氯芬酸钠片组大鼠炎症因子含量显著降低[(24.6±3.3)pg/ml,(151±21)pg/ml,(61±16)pg/ml]差异均有统计学意义(t=2.296,P<0.05;t=2.909,P<0.05;t=2.352,P<0.05)。和0.9%氯化钠注射液组比较,免疫组织化学法和蛋白质免疫印迹法检测的模型组踝关节NF-κBp65、MyD88、TLR4的蛋白表达显著增加;与模型组比较,双氯芬酸钠片组显著抑制踝关节组织的NF-κBp65、MyD88、Objective Sodium urate was used to induce acute gouty arthritis rat model,and to observe the inflammatory response of rats and the intervention effect of diclofenac sodium on the expression of Toll-like receptor-related(TLR)protein of ankle joint.Methods Thirty males specific pathogen free(SPF)grade Wistar rats were used to develop the models.Random number table method was used to divide the rats into normal saline control group,model group,and drug group(diclofenac sodium t 1.35 mg/g body weight),10 rats in each group.After fully grinding the sodium urate crystals,an appropriate amount of saline and Tween-80(9∶1)was added to make a suspension,and the sodium urate crystals(25 mg/ml)were injected to the right posterior ankle of the rats in the model and drug groups.The solution was 0.2 ml,and rats in the sham group were injected with 0.2 ml of normal saline at the same location.After the model was established,drug and equal volume of purified water were administrated intragastrically once a day for 7 days.The toe volume device was used to measure the joint swelling of the rat(at 4 h,8 h,24 h,48 h,72 h),and blood was taken from the abdominal aorta after anesthesia to determine the rat kidney function,interleukin(IL)-1β,IL-6 and tumor necrosis factor-α(TNF-α)content,the rat ankle joint TLR4,myeloid differentiation factor(MyD88),NF-κBp65 protein expression were determined using Western blot and immunohistochemical methods.Multiple comparisons were carried out using single factor analysis of variance(ANOVA),comparing the two groups by using LSD-t,the comparison of different times using repetitive measure analysis of variance(repeated measures).Results After the models were established,the rat's right ankle joint showed various degrees of redness,slow walking,and unresponsiveness.Compared with the normal saline control group,under the light microscope,the ankle synovial cells of the model group proliferated,with localized degeneration and necrosis,and many inflammatory cell infiltration.The rat serum inflammatory

关 键 词：尿酸钠 急性痛风性关节炎 炎性反应 Toll样受体通路 双氯芬酸钠 动物模型 

分 类 号：R589.7[医药卫生—内分泌] R-332[医药卫生—内科学]